Detection of chromosome translocation products in single interphase cell nuclei

Aneuploidy involving chromosome 1 in failed-fertilized human oocytes is unrelated to maternal age

“…The probe specific for satellite II DNA of chromosome 16 was prepared from clone pHuR195 (32) and labeled with FITC as well as digoxigenin. The single copy DNA probe specific for chromosome 21 (YAC 141G6) was selected from the Genethon/CEPH yeast artificial chromosome (YAC) library (33) and labeled with FITC as previously described (34). The DNA probe specific for chromosome 18 (CEP18) labeled with Spectrum Aqua was obtained from the manufacturer (Vysis).…”

“…Between 1.0 and 3.0 µL of each probe along with 1 µL of human COT1 TM DNA (1 mg/mL, Invitrogen) and 1 µL salmon sperm DNA (10 mg/mL, 3 -5 , Boulder, CO) were precipitated with 1 µL glycogen (1 mg/mL, Roche Molecular Biochemicals) and one-tenth volume of 3 M sodium acetate in two volumes of 2-propanol. The pellet was air-dried and resuspended in 3 µL water, before addition of 7 µL of hybridization master mix (34,35). The total volume of hybridization mixture containing 55% formamide (FA) and 10% dextran sulphate in sodium citrate buffer (2X SSC, 0.3 M NaCl, 0.03 M Na 3 citrate·2H 2 O, pH 7.0) was 10 µL.…”

“…The digoxigenin-labeled probes were detected with anti-digoxigenin rhodamine-conjugated antibody (20 µg/mL, Roche Molecular Biochemicals) (34). The slides were then washed three times in 2X SSC for 10 min each at 20 • C. Finally, the slides were mounted in 8 µL of 4 ,6-diamidino-2-phenylindole [DAPI, 0.05 µg/mL in antifade solution (34), Calbiochem, La Jolla, CA] to counterstain the DNA.…”

BAC-FISH assays delineate complex chromosomal rearrangements in a case of post-Chernobyl childhood thyroid cancer.

“…As the chromosome 9-specific example in this paper shows, only a handful of overnight hybridizations are required to localize the breakpoint to a single pool and open the door to high resolution analysis using single BAC clones. Thus, the proposed BAC pooling strategy seems capable to provide breakpoint information as well as DNA probes suitable for metaphase or interphase cell analysis in only 2–3 weeks, a significant improvement over previous methods [44,45]. …”

Kinase Expression and Chromosomal Rearrangements in Papillary Thyroid Cancer Tissues: Investigations at the Molecular and Microscopic Levels

“…For more than a decade, our laboratories have been involved in the preparation of breakpoint-specific probes and assays (12, 37-40). Initially, we used probes prepared from yeast artificial chromosome (YAC) clones, and it took on average 8-10 week to prepare probes suitable for interphase analysis (37).…”